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Khalid Shahin, Jose Gustavo Ramirez-Paredes, Graham Harold, Benjamin Lopez-Jimena, Alexandra Adams, Manfred Weidmann
Francisella noatunensis subsp. orientalis (Fno) is the causative agent of piscine francisellosis in warm water fish including tilapia. The disease induces chronic granulomatous inflammation with high morbidity and can result in high mortality. Early and accurate detection of Fno is crucial to set appropriate outbreak control measures in tilapia farms. Laboratory detection of Fno mainly depends on bacterial culture and molecular techniques. Recombinase polymerase amplification (RPA) is a novel isothermal technology that has been widely used for the molecular diagnosis of various infectious diseases...
2018: PloS One
Guido Rocchigiani, Valentina Virginia Ebani, Simona Nardoni, Fabrizio Bertelloni, Alice Bascherini, Alessandro Leoni, Francesca Mancianti, Alessandro Poli
Data about the spreading of arthropod-borne pathogens among hare populations are very scant, so the aim of the present preliminary study was to investigate, through molecular analysis, the occurrence of Anaplasma phagocytophilum, Bartonella sp., Borrelia burgdorferi sensu lato, Coxiella burnetii, Francisella tularensis, Leishmania spp. and piroplasms DNA in blood of 51 wild hares (Lepus europaeus) living in protected areas in Tuscany. All hares resulted negative for A. phagocytophilum, Bartonella sp., B. burgdorferi s...
February 5, 2018: Infection, Genetics and Evolution
Ralph Anthony Stidham, David B Freeman, Robert L von Tersch, Peter J Sullivan, Samantha D Tostenson
Introduction: Tularemia is a rare but potentially fatal disease that develops in numerous wild and domestic animals, including lagomorphs, rodents, cats, and humans.  Francisella tularensis bacterium, the causative agent of tularemia, was identified by veterinary personnel at Fort Riley, Kansas during a routine post-mortum evaluation of a domestic feline. However, before formal diagnosis was confirmed, the sample was sent and prepared for rabies testing at the Department of Defense (DoD) U...
January 18, 2018: PLoS Currents
Adva Mechaly, Einat Vitner, Haim Levy, Shay Weiss, Elad Bar-David, David Gur, Michal Koren, Hila Cohen, Ofer Cohen, Emanuelle Mamroud, Morly Fisher
Multiplexed detection technologies are becoming increasingly important given the possibility of bioterrorism attacks, where the range of suspected pathogens can vary considerably. In this work, we describe the use of Luminex MagPlex magnetic microspheres for the construction of two multiplexed diagnostic suspension arrays, enabling the antibody-based detection of bacterial pathogens and their related disease biomarkers directly from blood cultures.The first 4-plex diagnostic array enabled the detection of both anthrax and plague infections using soluble disease biomarkers, including PA (Protective Antigen) and anthrax capsular antigen for anthrax detection and the capsular F1 and LcrV antigens for plague detection...
January 31, 2018: Journal of Clinical Microbiology
Jonathan G Gerhart, H Auguste Dutcher, Amanda E Brenner, Abraham S Moses, Libor Grubhoffer, Rahul Raghavan
Bacterial endosymbionts of ticks are of interest due to their close evolutionary relationships with tick-vectored pathogens. For instance, while many ticks contain Francisella-like endosymbionts (FLEs), others transmit the mammalian pathogen Francisella tularensis. We recently sequenced the genome of an FLE present in the hard tick Amblyomma maculatum (FLE-Am) and showed that it likely evolved from a pathogenic ancestor. In order to expand our understanding of FLEs, in the current study we sequenced the genome of an FLE in the soft tick Ornithodoros moubata (FLE-Om) and compared it to the genomes of FLE-Am, F...
January 29, 2018: Genome Biology and Evolution
Alla Zamyatina
The immediate immune response to infection by Gram-negative bacteria depends on the structure of a lipopolysaccharide (LPS, also known as endotoxin), a complex glycolipid constituting the outer leaflet of the bacterial outer membrane. Recognition of picomolar quantities of pathogenic LPS by the germ-line encoded Toll-like Receptor 4 (TLR4) complex triggers the intracellular pro-inflammatory signaling cascade leading to the expression of cytokines, chemokines, prostaglandins and reactive oxygen species which manifest an acute inflammatory response to infection...
2018: Beilstein Journal of Organic Chemistry
Maha Alqahtani, Zhuo Ma, Harshada Ketkar, Ragavan Varadharajan Suresh, Meenakshi Malik, Chandra Shekhar Bakshi
Francisella tularensis; the causative agent of tularemia lacks typical bacterial virulence factors and toxins, but still exhibits extreme virulence. The bacterial multi-drug efflux systems consist of an inner membrane, a transmembrane membrane fusion protein and an outer membrane (OM) component that form a contiguous channel for secretion of multitude of bacterial products. Francisella contains three orthologs of the OM proteins; two of these termed as TolC and FtlC are important for tularemia pathogenesis...
January 29, 2018: Journal of Bacteriology
Sivakumar Periasamy, Jonathan A Harton
Bacterial pneumonia is a common risk factor for acute lung injury and sepsis-mediated death, but the mechanisms underlying the overt inflammation and accompanying pathology are unclear. Infiltration of immature myeloid cells (IMC) and necrotizing inflammation mediate severe pathology and death during pulmonary infection with Francisella tularensis. However, eliciting mature myeloid cells provides protection. Yet, the host factors responsible for this pathologic IMC response are unknown. Here, we report that while the influx of both mature and immature is strictly MyD88-dependent, the IL-1R receptor mediates an important dual function via its ligands IL-1α and IL-1β...
January 24, 2018: Journal of Infectious Diseases
N J Mank, S Pote, K A Majorek, A K Arnette, V G Klapper, B K Hurlburt, M Chruszcz
Aspartate β-semialdehyde dehydrogenase (ASADH) is an enzyme involved in the diaminopimelate pathway of lysine biosynthesis. It is essential for the viability of many pathogenic bacteria and therefore has been the subject of considerable research for the generation of novel antibiotic compounds. This manuscript describes the first structure of ASADH from Francisella tularensis, the causative agent of tularemia and a potential bioterrorism agent. The structure was determined at 2.45 Å resolution and has a similar biological assembly to other bacterial homologs...
January 1, 2018: Acta Crystallographica. Section F, Structural Biology Communications
Liliya M Pekova, Magdalena P Baymakova, Pavlina O Parousheva, Mariya D Fartunova, Nikolay E Dimitrov, Iskra A Tomova
We present here the first five human cases with tularemia from two regions in South Bulgaria in which there had been no previous report of the infection. The cases occurred over a period of 8 months (December 2014 - August 2015). They were treated at the Department of Infectious Diseases in Stara Zagora University Hospital, Bulgaria. We present the clinical, epidemiological and laboratory data for four men and one woman (age range 52 to 73 years). Three men were hunters, four patients took part in handling, preparing/skinning and cooking the game animals...
December 20, 2017: Folia Medica
Brice Lagrange, Sacha Benaoudia, Pierre Wallet, Flora Magnotti, Angelina Provost, Fanny Michal, Amandine Martin, Flaviana Di Lorenzo, Bénédicte F Py, Antonio Molinaro, Thomas Henry
Caspase-4/5 in humans and caspase-11 in mice bind hexa-acylated lipid A, the lipid moeity of lipopolysaccharide (LPS), to induce the activation of non-canonical inflammasome. Pathogens such as Francisella novicida express an under-acylated lipid A and escape caspase-11 recognition in mice. Here, we show that caspase-4 drives inflammasome responses to F. novicida infection in human macrophages. Caspase-4 triggers F. novicida-mediated, gasdermin D-dependent pyroptosis and activates the NLRP3 inflammasome. Inflammasome activation could be recapitulated by transfection of under-acylated LPS from different bacterial species or synthetic tetra-acylated lipid A into cytosol of human macrophage...
January 16, 2018: Nature Communications
L M Matz, K Y Kamdar, M E Holder, G A Metcalf, G M Weissenberger, Q Meng, V Vee, Y Han, D M Muzny, R A Gibbs, C L Johnson, P A Revell, J F Petrosino
The accumulation of sequenced Francisella strains has made it increasingly apparent that the 16S rRNA gene alone is not enough to stratify the Francisella genus into precise and clinically useful classifications. Continued whole-genome sequencing of isolates will provide a larger base of knowledge for targeted approaches with broad applicability. Additionally, examination of genomic information on a case-by-case basis will help resolve outstanding questions regarding strain stratification. We report the complete genome sequence of a clinical isolate, designated here as F...
December 6, 2017: Diagnostic Microbiology and Infectious Disease
Tong Zhang, Qiufeng Zheng, Xin Yi, Hong An, Yaling Zhao, Siqi Ma, Guohui Zhou
Recently, CRISPR-Cas (clustered, regularly interspaced short palindromic repeats-CRISPR associated proteins) system has been used to produce plants resistant to DNA virus infections. However, there is no RNA virus control method in plants that uses CRISPR-Cas system to target the viral genome directly. Here we reprogrammed the CRISPR-Cas9 system from Francisella novicida to confer molecular immunity against RNA viruses in Nicotiana benthamiana and Arabidopsis plants. Plants expressing FnCas9 and sgRNA specific for the cucumber mosaic virus (CMV) or tobacco mosaic virus (TMV) exhibited significantly attenuated virus infection symptoms and reduced viral RNA accumulation...
January 11, 2018: Plant Biotechnology Journal
Donald J Steiner, Yoichi Furuya, Dennis W Metzger
Opsonizing antibody is a critical component of the host protective immune response against many respiratory pathogens. However, the role of antibodies in protection against pulmonary infection with highly virulent Francisella tularensis strain SchuS4 is unclear and the mechanism that allows F. tularensis to evade antibody-mediated bacterial clearance is not fully understood. We have now found that depletion of alveolar macrophages reveals an otherwise cryptic protective effect of opsonizing antibody. While antibody opsonization alone failed to confer any survival benefit against SchuS4 lung infection, significant protection was observed when mice were depleted of alveolar macrophages prior to infection...
January 8, 2018: Infection and Immunity
Phillip M Balzano, Aimee L Cunningham, Christen Grassel, Eileen M Barry
Francisella tularensis is a Gram negative facultative intracellular coccobacillus that can infect a wide variety of hosts. In humans, F. tularensis causes the zoonosis tularemia following insect bites, ingestion, inhalation, and the handling of infected animals. That a very small inoculum delivered by the aerosol route can cause severe disease, coupled with the possibility of its use as an aerosolized bioweapon, have led to the classification of Francisella tularensis as a Category A select agent and has renewed interest in the formulation of a vaccine...
January 8, 2018: Infection and Immunity
Osman Salih, Shaoda He, Sara Planamente, Lasse Stach, James T MacDonald, Eleni Manoli, Sjors H W Scheres, Alain Filloux, Paul S Freemont
Pseudomonas aeruginosa has three type VI secretion systems (T6SSs), H1-, H2-, and H3-T6SS, each belonging to a distinct group. The two T6SS components, TssB/VipA and TssC/VipB, assemble to form tubules that conserve structural/functional homology with tail sheaths of contractile bacteriophages and pyocins. Here, we used cryoelectron microscopy to solve the structure of the H1-T6SS P. aeruginosa TssB1C1 sheath at 3.3 Å resolution. Our structure allowed us to resolve some features of the T6SS sheath that were not resolved in the Vibrio cholerae VipAB and Francisella tularensis IglAB structures...
December 21, 2017: Structure
David E Place, David R Williamson, Yevgeniy Yuzefpolskiy, Bhuvana Katkere, Surojit Sarkar, Vandana Kalia, Girish S Kirimanjeswara
Progress towards a safe and effective vaccine for the prevention of tularemia has been hindered by a lack of knowledge regarding the correlates of protective adaptive immunity and a lack of tools to generate this knowledge. CD8+ T cells are essential for protective immunity against virulent strains of Francisella tularensis, but to-date, it has not been possible to study these cells in an antigen-specific manner. Here, we report the development of a tool for expression of the model antigen ovalbumin (OVA) in F...
2017: PloS One
Ramya Sundaresan, Hari Priya Parameshwaran, S D Yogesha, Mark Walter Keilbarth, Rakhi Rajan
CRISPR-Cas systems provide bacteria and archaea with sequence-specific protection against invading mobile genetic elements. In the presence of divalent metal ions, Cas9 and Cas12a (formerly Cpf1) proteins target and cleave DNA that is complementary to a cognate guide RNA. The recognition of a protospacer adjacent motif (PAM) sequence in the target DNA by Cas9 and Cas12a is essential for cleavage. This RNA-guided DNA targeting is widely used for gene-editing methods. Here, we show that Francisella tularensis novicida (Fno) Cas12a, FnoCas9, and Streptococcus pyogenes Cas9 (SpyCas9) cleave DNA without a guide RNA in the presence of Mn2+ ions...
December 26, 2017: Cell Reports
Artem Rogovskyy, Maliha Batool, David C Gillis, Patricia J Holman, Igor V Nebogatkin, Yuliya V Rogovska, Maksym S Rogovskyy
Lyme borreliosis (LB) is caused by tick-borne spirochetes of the Borrelia burgdorferi sensu lato complex. LB is the most prevalent vector-borne illness in Ukraine, but current data on the prevalence of LB pathogens in their tick vector, Ixodes ricinus, are lacking. I. ricinus ticks may also carry Borrelia miyamotoi, an emerging relapsing fever group spirochete that has been implicated in human illness. Despite its zoonotic potential, the prevalence of B. miyamotoi in ticks has not been examined in Ukraine. Similarly, data on the prevalence of other important tick-borne pathogens, Anaplasma phagocytophilum, Babesia spp...
December 15, 2017: Ticks and Tick-borne Diseases
Amrita Lama, Samuel L Drennan, Rudd C Johnson, Grace L Rubenstein, Eric D Cambronne
It is established that the human pathogen Legionella pneumophila becomes significantly augmented for infection of macrophages after intracellular growth in amoebae when compared to like-strains cultivated in laboratory media. Based on this observation, we reasoned that the most critical virulence determinants of L.p. are expressed by responding to stimuli generated by the protozoan host specifically; a process we term "protozoan-priming." We sought to identify L.p. virulence factors that were required for replication in amoebae in order to highlight the genes necessary for production of the most infectious form of the bacterium...
2017: Frontiers in Cellular and Infection Microbiology
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